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Feb 28, 1996 - These contain the cuboidal [Fe3S4]0 fragment with S = 2 which is “spin-isolated” in the cluster because the remaining metal site is...
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J. Am. Chem. Soc. 1996, 118, 1966-1980

The Cuboidal Fe3S4 Cluster: Synthesis, Stability, and Geometric and Electronic Structures in a Non-Protein Environment Jian Zhou,† Zhengguo Hu,‡ Eckard Mu1 nck,*,‡ and R. H. Holm*,† Contribution from the Departments of Chemistry, HarVard UniVersity, Cambridge, Massachusetts 02138, and Carnegie-Mellon UniVersity, Pittsburgh, PennsylVania 15213 ReceiVed NoVember 10, 1995X

Abstract: Of the three known low-nuclearity iron-sulfur clusters in metallobiomolecules with the core units Fe2S2, Fe4S4, and Fe3S4, the last has not been obtained in stable form outside a protein environment. We describe a direct route to such clusters in the [Fe3S4]0 oxidation state, and demonstrate an effective stereochemical and electronic structural congruence with the native cluster. The synthesis is based on iron-site-differentiated clusters. Reaction of [Fe4S4(LS3)(SEt)]2- with (Et3NH)(OTf) affords [Fe4S4(LS3)(OTf)]2-, whose unique site is activated toward terminal ligand substitution. Treatment with 1 equiv of (Et4N)2(Meida) affords [Fe4S4(LS3)(Meida)]3-, which is readily converted to [Fe3S4(LS3)]3- with 1-2 equiv of additional reactant. The trinuclear cluster is formed by abstraction of Fe2+ from a precursor cubane-type [Fe4S4]2+ core and complexation as [Fe(Meida)2]2-. An analogous procedure starting with [Fe4Se4(LS3)(SEt)]2- yields [Fe3Se4(LS3)]3-. The compound (Et4N)3[Fe3S4(LS3)]‚MeCN crystallizes in orthorhombic space group P212121 with no imposed symmetry. An X-ray structure solution demonstrates the presence of the desired cuboidal [Fe3(µ3-S)(µ2-S)3]0 core in a complex of absolute configuration ∆. Property comparisons support the cuboidal structure for [Fe3Se4(LS3)]3-. A series of reactions in the systems [Fe4S4(SEt)4]2-/ (Et4N)2(Meida) and [Fe3S4(LS3)]3-/NaSEt in Me2SO disclose that, while cuboidal [Fe3S4(SEt)3]3- is formed in both systems, it is one of several cluster products and tends to decay with time. [Fe3S4(LS3)]3- is completely stable in anaerobic solutions at ambient temperature. Consequently, the semirigid cavitand ligand LS3 is conspicuously superior to a simple monodentate thiolate in stabilizing the [Fe3S4]0 core. The cuboidal core is metrically very similar in structure to the cubane core of [Fe4S4(LS3)Cl]2- and to protein-bound Fe3S4 clusters. Voltammetry of [Fe3S4(LS3)]3reveals a reversible three-membered electron transfer series which includes the core states [Fe3S4]1+,0,1-. The electronic structures of [Fe3S4(LS3)]3- and [Fe3Se4(LS3)]3- were investigated by Mo¨ssbauer and EPR spectroscopies. These studies reveal that the synthetic clusters, like the protein-bound clusters, have an electronic ground state with cluster spin S ) 2 that arises from an interplay of Heisenberg and double exchange between the sites of a delocalized Fe2+Fe3+ pair and an Fe3+ site. The zero-field splittings of the S ) 2 multiplet and the entire set of 57Fe hyperfine parameters of the synthetic clusters match those of the protein-bound clusters. Evidently, protein structure is not required to sustain the cuboidal geometry nor the spin-quintet ground state and its attendant electron distribution and magnetic interactions. We conclude that the clusters [Fe3Q4(LS3)]3- (Q ) S, Se) are accurate structural and electronic analogues of the cuboidal sites in native and selenide-reconstituted proteins. No cluster containing a discrete cuboidal Fe3S4 core has previously been isolated in substance.

Introduction1 The pervasive occurrence in biology of clusters 1 containing the cubane-type Fe4S4 core is widely recognized. It has become increasingly evident within the last decade that the structurally derived clusters 2 having the cuboidal Fe3S4 core are also extensively distributed. Furthermore, this cluster type is intrinsic to at least some proteins rather than necessarily being an artifact arising from oxidative damage of native clusters 1. Schematic structures of 1 and 2 are set out in Figure 1. The first 3-Fe †

Harvard University. Carnegie Mellon University. X Abstract published in AdVance ACS Abstracts, February 15, 1996. (1) Abbreviations: Ac, Azotobacter chroococcum; AV, Azotobacter Vinelandii; bpy, 2,2′-bipyridyl; CV, Chromatium Vinosum; Da, DesulfoVibrio africanus; Dg, DesulfoVibrio gigas, Fd, ferredoxin; LS3, 1,3,5-tris((4,6dimethyl-3-mercaptophenyl)thio)-2,4,6-tris(p-tolylthio)benzene(3-); Meida, N-methylimidodiacetate; Me5dien, 1,1,4,7,7-pentamethyldiethylenetriamine; Me6tren, tris(2-(dimethylamino)ethyl)amine; NHE, normal hydrogen electrode; OTf, triflate; Pf, Pyrococcus furiosus; Q, S or Se; SCE, standard calomel electrode; Smes, mesitylthiolate(1-); Stibt, 2,4,6-triisopropylbenzenethiolate(1-); tacn, 1,4,7-triazacyclononane. ‡

0002-7863/96/1518-1966$12.00/0

clusters were recognized by Mu¨nck and co-workers,2 who found from quantitative EPR and Mo¨ssbauer measurements one spin associated with three iron atoms in one molecule of AV Fd I and Dg Fd II.2 Accounts of early developments in the area are available.3 Subsequently, these clusters have been extensively characterized by magnetic and spectroscopic means, and structure 2 established crystallographically for inactive aconitase,4 Dg Fd II,5 and AV Fd I6 in their oxidized ([Fe3S4]1+) states. AV Fd I is representative of 7-Fe proteins which contain one (2) (a) Emptage, M. H.; Kent, T. A.; Huynh, B. H.; Rawlings, J.; OrmeJohnson, W. H.; Mu¨nck, E. J. Biol. Chem. 1980, 255, 1793. (b) Huynh, B. H.; Moura, J. J. G.; Moura, I.; Kent, T. A.; LeGall, J.; Xavier, A. V.; Mu¨nck, E. J. Biol. Chem. 1980, 255, 3242. (3) At this stage (1980-1983), the clusters were usually formulated as Fe3S3: (a) Beinert, H.; Thomson, A. J. Arch. Biochem. Biophys. 1983, 222, 333. (b) Mu¨nck, E. In Iron-Sulfur Proteins; Spiro, T. G., Ed.; WileyInterscience: New York, 1982; Chapter 4. (4) Robbins, A. H.; Stout, C. D. Proteins 1989, 5, 289. (5) (a) Kissinger, C. R.; Adman, E. T.; Sieker, L. C.; Jensen, L. H. J. Am. Chem. Soc. 1988, 110, 8721. (b) Kissinger, C. R.; Sieker, L. C.; Adman, E. T.; Jensen, L. H. J. Mol. Biol. 1991, 219, 693. (6) (a) Stout, C. D. J. Mol. Biol. 1989, 205, 545. (b) Soman, J.; Iismaa, S.; Stout, C. D. J. Biol. Chem. 1991, 266, 21558. (c) Stout, C. D. J. Biol. Chem. 1993, 268, 25920.

© 1996 American Chemical Society

The Cuboidal Fe3S4 Cluster

Figure 1. Summary of the oxidation and spin states and the interconversion of Fe4S4 and Fe3S4 clusters. Terminal ligand L is H2O/ HO- or a non-cysteinyl protein group. Reported potential limits (vs NHE) for two redox steps are indicated; the single value of the [Fe4S4]3+/2+ couple is for aconitase.29 Commonly used redox reagents are shown; dithionite cannot fully reduce [Fe4S4]2+/1+ couples with the lower potentials. Allowance is made for protonation of [Fe3S4]0,2clusters, but the number of bound protons is uncertain. The all-ferrous [Fe3S4]2- core is postulated, not proven.

Fe3S4 and one Fe4S4 cluster. At least in smaller proteins (j10 kDa), the appearance of an Fe3S4 cluster can be correlated with departure from the cysteinyl triplet amino acid sequence CysX-X-Cys-X-X-Cys common to the vast majority of proteins which incorporate Fe4S4 clusters.7 In such cases, two Cys residues function as cluster ligands; the remaining Cys ligand of the trinuclear cluster is derived from a residue considerably separated along the chain from the sequence run. For example, in Da Fd III,8 the central Cys residue is replaced by Asp,9 such that the Fe3S4 center is ligated by Cys-11/17/51. When the protein is converted to the 8-Fe form, one Fe4S4 cluster retains these three ligands; the remaining Fe site is bound by H2O/ HO- or possibly the carboxylate group of Asp-14.10 In the 4-Fe protein Pf Fd, the central Cys position is also occupied by Asp, which remains ligated to the cluster in all oxidation states.11 While this protein does not lose iron during purification, it is readily converted to the Fe3S4 form by oxidation.12 The study of protein-bound Fe3S4 clusters has had a major impact on the magnetochemistry of Fe-S clusters and other exchange-coupled systems. In the reduced state, designated [Fe3S4]0, the cluster formally contains two Fe3+ and one Fe2+. However, Mo¨ssbauer studies of AV Fd I and Dg Fe II showed that the S ) 2 cluster ground state consists of a valencedelocalized pair of iron atoms with mean oxidation state Fe2.5+ and one valence-trapped Fe3+ site.2 The presence of a delocalized Fe2+Fe3+ pair requires a spin coupling model that takes explicitly into account the delocalization. This problem was solved by introduction of a new spin Hamiltonian that includes (7) Holm, R. H. AdV. Inorg. Chem. 1992, 38, 1. (8) (a) Armstrong, F. A.; George, S. J.; Cammack, R.; Hatchikian, E. C.; Thomson, A. J. Biochem. J. 1989, 264, 265. (b) George, S. J.; Armstrong, F. A.; Hatchikian, E. C.; Thomson, A. J. Biochem. J. 1989, 264, 275. (9) Bovier-Lapierre, G.; Bruschi, M.; Bonicel, J.; Hatchikian, E. C. Biochim. Biophys. Acta 1987, 913, 20. (10) Butt, J. N.; Sucheta, A.; Armstrong, F. A.; Breton, J.; Thomson, A. J.; Hatchikian, E. C. J. Am. Chem. Soc. 1993, 115, 1413. (11) Calzolai, L.; Gorst, C. M.; Zhao, Z.-H.; Teng, Q.; Adams, M. W. W.; La Mar, G. N. Biochemistry 1995, 34, 11373. (12) (a) Aono, S.; Bryant, F. O.; Adams, M. W. W. J. Bacteriol. 1989, 171, 3433. (b) Conover, R. C.; Kowal, A. T.; Fu, W.; Park, J.-B.; Aono, S.; Adams, M. W. W.; Johnson, M. K. J. Biol. Chem. 1990, 265, 8533.

J. Am. Chem. Soc., Vol. 118, No. 8, 1996 1967 a resonance delocalization term (double exchange) in addition to the familiar Heisenberg (JSi‚Sj) interaction.13 Inclusion of double exchange led to a picture where the observed S ) 2 spin of the ground state arises from antiparallel alignment of S ) 5/2 spin of the Fe3+ site with the S ) 9/2 spin of the delocalized pair; the S ) 9/2 spin results from parallel alignment of the core spins of the delocalized pair by the itinerant electron.13 The concept of double exchange, introduced by Zener14 and further developed by Anderson and Hasegawa,15 has proven fruitful for the description of the magnetic properties of [Fe3S4]0 clusters13,16-19 and [Fe4S4]3+/2+/1+ clusters,16,18-21 as well as the coupled siroheme-Fe4S4 chromophore of E. coli sulfite reductase.22 Experimental values of the double exchange parameter, B, have not been reported for any Fe-S cluster.23 However, broken symmetry density functional calculations by Noodleman and co-workers19 suggest that the double exchange contribution is substantial; namely, B ≈ 700 cm-1 for the [Fe4S4] clusters and B ≈ 400 cm-1 for [Fe3S4]0 clusters. It has been pointed out25 that an S ) 2 ground state could also arise from weak double exchange if different J values for the three Fe pairs were considered. On the other hand, Borshch et al.17 have shown that a combination of Vibronic interactions and double exchange, even in the absence of Heisenberg-Direc-van Vleck exchange, leads to an S ) 2 ground state consisting of a delocalized Fe2+Fe3+ pair and a trapped Fe3+ site. Most recently, Bominaar et al.22 have proposed a novel vibronic term that arises from the distance dependence of the double exchange interaction. Clearly, the combination of vibronic interactions with HDvV exchange and double exchange provides a rich new realm of magnetochemical phenomena. Shortly after the discovery of 3-Fe clusters in AV Fd I and Dg Fd II, the new cluster type was shown to be present in inactive aconitase. However, when this enzyme was activated by addition of labeled Fe2+, the added 57Fe was found to be incorporated into a subsite of an Fe4S4 cluster, demonstrating that the 3-Fe cluster could be converted into the familiar cubane.26a This conversion, soon thereafter confirmed and elaborated for Dg Fd II,27 provided a strong hint that the 3-Fe cluster had an Fe3S4 core.3 The observation of reversible Fe3S4T Fe4S4 conversions in aconitase and Dg Fd II, where three iron sites are protected by the protein matrix, suggested a (13) Papaefthymiou, V.; Girerd, J.-J.; Moura, I.; Moura, J. J. G.; Mu¨nck, E. J. Am. Chem. Soc. 1987, 109, 4703. (14) Zener, C. Phys. ReV. 1951, 82, 403. (15) Anderson, P. W.; Hasegawa, H. Phys. ReV. 1955, 100, 675. (16) Sontum, S. F.; Noodleman, L.; Case, D. A. In The Challenge of d and f Electrons; Salahub, D. R., Zerner, M., Eds.; ACS Symposium Series 394; American Chemical Society: Washington, DC, 1998; Chapter 26. (17) Borshch, S. A.; Bominaar, E. L.; Blondin, G.; Girerd, J.-J. J. Am. Chem. Soc. 1993, 115, 5155. (18) Bominaar, E. L.; Borshch, S. A.; Girerd, J.-J. J. Am. Chem. Soc. 1994, 116, 5362. (19) Noodleman, L.; Case, D. A. AdV. Inorg. Chem. 1992, 38, 423. (20) Noodleman, L.; Peng, C. Y.; Case, D. A.; Mouesca, J.-M. Coord. Chem. ReV. 1994, 144, 199. (21) Belinskii, M. Chem. Phys. 1993, 173, 27. (22) Bominaar, E. L.; Hu, Z.; Mu¨nck, E.; Girerd, J.-J.; Borshch, S. J. Am. Chem. Soc. 1995, 117, 6976. (23) To date, [Fe3S4]0 is the Fe-S cluster with the smallest nuclearity that exhibits valence delocalization. However, an S ) 9/2 spin state for an [Fe2S2]1+ cluster has recently been reported.24 Mo¨ssbauer spectroscopy is ideally suited to assess whether the cluster consists of a delocalized Fe2+Fe3+ pair; such studies, however, have not yet been reported. (24) Crouse, B. R.; Meyer, J.; Johnson, M. K. J. Am. Chem. Soc. 1995, 117, 9612. (25) Bertini, I.; Briganti, F.; Luchinat, C. Inorg. Chim. Acta 1990, 175, 9. (26) (a) Kent, T. A.; Dreyer, J.-L.; Kennedy, M. C.; Huynh, B. H.; Emptage, M. H.; Beinert, H.; Mu¨nck, E. Proc. Natl. Acad. Sci. U.S.A. 1982, 79, 1096. (b) Emptage, M. H.; Dreyer, J.-L.; Kennedy, M. C.; Beinert, H. J. Biol. Chem. 1983, 258, 11106. (c) Kent, T. A.; Emptage, M. H.; Merkle, H.; Kennedy, M. C.; Beinert, H.; Mu¨nck, E. J. Biol. Chem. 1985, 260, 6871.

1968 J. Am. Chem. Soc., Vol. 118, No. 8, 1996 strategy for synthesis of the cubane Fe3S4 core that was based on iron-site-differentiated Fe4S4 clusters. Summarized in Figure 1 are the known oxidation and spin ground states of protein-bound Fe4S4 and Fe3S4 clusters, ranges of redox potentials, and interconversion reactions of clusters and several common reactants which effect them. The upper portion of the scheme, involving the interconversion of clusters 1 and 2, is drawn largely from the extensive study of aconitase by Beinert, Kennedy, Mu¨nck, and their co-workers.7,26,28 Redox potentials are those of aconitase29 and, primarily, 7-Fe proteins8a,30 which exhibit the couples [Fe4S4]2+/1+ and [Fe3S4]1+/0; ranges of values are indicated. Chemical oxidation of the [Fe4S4]2+ state in a variety of proteins yields the [Fe3S4]1+ (S ) 1/2) cluster, identified by characteristic MCD and Mo¨ssbauer spectra and an EPR signal centered at g ∼ 2.01. This transformation may proceed by initial formation of the highly oxidized state [Fe4S4]3+ (normally stable only in the “high-potential” subclass of ferredoxins), an Fe3S4 fragment of which is insufficiently electron rich to retain Fe2+. This ion is released and stabilized in solvated or chelated form, affording the oxidized [Fe3S4]1+ cluster. Indeed, Tong and Feinberg,29 using direct square-wave voltammetry of aconitase, report detection of the [Fe4S4]3+/2+ couple at +100 mV and the subsequent spontaneous reaction [Fe4S4]3+ f [Fe3S4]1+ + Fe2+. With entry to the Fe3S4 cluster regime by oxidative cluster degradation, it has been shown that the [Fe3S4]1+ state of Da Fd III has no detectable affinity to bind Fe2+ and other divalent ions.31 Dithionite reduction affords the [Fe3S4]0 state. The pH dependence of [Fe3S4]1+/0 potentials and MCD spectra indicate that one proton is transferred upon reduction in certain cases (aconitase,29 AV Fd I,30ab Ac Fd I,30c) but not another (Da Fd III8a). All Fe3S4 proteins exhibit a second redox process at about -800 mV. In the case of Da Fd III, which is the most thoroughly studied,32 the reduction involves the addition of two electrons and the net uptake of as many as three protons.33 In Figure 1, we assume for accounting purposes that the reduction product is the all-ferrous [Fe3S4]2- cluster, and for it and its precursor we allow for the possibility that the clusters are protonated. Many of the electrochemical features of proteinbound Fe3S4 clusters have been summarized by Armstrong.33 Whereas analogues of native Fe2S2 and Fe4S4 clusters are readily prepared,34,35 extensive attempts in this laboratory to (27) Moura, J. J. G.; Moura, I.; Kent, T. A.; Lipscomb, J. D.; Huynh, B. H.; LeGall, J.; Xavier, A. V.; Mu¨nck, E. J. Biol. Chem. 1982, 257, 6259. (28) (a) Emptage, M. H. In Metal Clusters in Proteins; Que, L., Jr., Ed.; ACS Symposium Series 372; American Chemical Society: Washington, DC, 1988; Chapter 17. (b) Kennedy, M. C.; Stout, C. D. AdV. Inorg. Chem. 1992, 38, 323. (29) Tong, J.; Feinberg, B. A. J. Biol. Chem. 1994, 269, 24920. (30) (a) Iismaa, S. E.; Va`zquez, A. E.; Jensen, G. M.; Stephens, P. J.; Butt, J. N.; Armstrong, F. A.; Burgess, B. K. J. Biol. Chem. 1991, 266, 21563. (b) Shen, B.; Martin, L. L.; Butt, J. N.; Armstrong, F. A.; Stout, C. D.; Jensen, G. M.; Stephens, P. J.; La Mar, G. N.; Gorst, C. M.; Burgess, B. K. J. Biol. Chem. 1993, 268, 25928. (c) Armstrong, F. A.; George, S. J.; Thomson, A. J.; Yates, M. G. FEBS Lett. 1988, 234, 107. (d) Ohnishi, T.; Blum, H.; Sato, S.; Nakazawa, K.; Hon-nami, K.; Oshima, T. J. Biol. Chem. 1980, 255, 345. (e) Iwasai, T.; Wakagi, T.; Isogai, Y.; Tanaka, K.; Iizuka, T.; Oshima, T. J. Biol. Chem. 1994, 269, 29444. (f) Teixeira, M.; Batista, R.; Campos, A. P.; Gomes, C.; Mendes, J.; Pacheco, I.; Anemuller, S.; Hagen, W. R. Eur. J. Biochem. 1995, 227, 322. (g) Cammack, R.; Rao, K. K.; Hall, D. O.; Moura, J. J. G.; Xavier, A. V.; Bruschi, M.; LeGall, J.; Deville, A.; Gayda, J. P. Biochim. Biophys. Acta 1977, 490, 311. (h) Moreno, C.; Macedo, A. L.; Moura, I.; LeGall, J.; Moura, J. J. G. J. Inorg. Biochem. 1994, 53, 219. (i) Battistuzzi, G.; Borsari, M.; Ferretti, S.; Luchinat, C.; Sola, M. Arch. Biochem. Biophys. 1995, 320, 149. (31) (a) Butt, J. N.; Armstrong, F. A.; Breton, J.; George, S. J.; Thomson, A. J.; Hatchikian, E. C. J. Am. Chem. Soc. 1991, 113, 6663. (b) Thomson, A. J.; Breton, J.; Butt, J. N.; Hatchikian, E. C.; Armstrong, F. A. J. Inorg. Biochem. 1992, 47, 197. (32) Armstrong, F. A.; Butt, J. N.; George, S. J.; Hatchikian, E. C.; Thomson, A. J. FEBS Lett. 1989, 259, 15. (33) Armstrong, F. A. AdV. Inorg. Chem. 1992, 38, 117.

Zhou et al. synthesize cuboidal Fe3S4 have failed. This core has been prepared, but in its isomeric linear [Fe3(µ2-S)4]1+ configuration.35 These observations and certain of the results in Figure 1 raise some fundamental questions as to the intrinsic properties of the Fe3S4 cluster, among which are the following. (i) Are there synthetic routes to the cluster other than oxidative degradation of an Fe4S4 cluster? (ii) Does cuboidal stereochemistry require stabilization by a protein matrix or a (thiolate) ligand of special design? (iii) Are the metric features of an Fe3S4 cluster closely similar to an Fe4S4 “parent” cluster (as protein results indicate4-6), or does the cluster relax to significantly different dimensions when, in effect, one iron atom is removed from the parent? (iv) Can the [Fe3S4]1- state, as yet undetected in proteins (unless stabilized by binding to a metal cation30h,31a,36,37), be attained? And can the all-ferrous [Fe3S4]2state be stabilized with, or without, protonation? (v) Is the potential order E([Fe4S4]2+/1+) j E([Fe3S4]1+/0) inherent or dependent on protein environment? (vi) Are the fine details of electronic structure (charge distribution, zero-field splitting, 57Fe quadrupole splittings, and hyperfine couplings) dependent on protein structure and its inherent asymmetry? (vii) Will the oxidation states [Fe3S4]0,1- bind metal ions and thus offer a general route to heterometal MFe3S4 clusters 3 (Figure 1)? No such route to MFe3S4 clusters currently exists, these clusters having been prepared by the seemingly disparate methods of self-assembly and reductive rearrangement of linear [Fe3S4]1+ clusters.7,38 Concerning point (i) above, we have recently devised a simple synthetic entry to [Fe3S4]0 clusters39 which we describe in detail here together with structure proof of the cuboidal cluster product. Also, we address points (ii)-(vi), leaving (vii) and the second part of (iv) for subsequent reports. Noting that apo-aconitase can be reconstituted to an active ([Fe4Se4]2+) form with an iron salt and selenide, and converted to an inactive ([Fe3Se4]1+) form by reaction with ferricyanide,40 we have included the preparation and characterization of a [Fe3Se4]0 cluster in this investigation. Experimental Section Preparation of Compounds. All operations were performed under a pure dinitrogen atmosphere using standard glovebox or Schlenk techniques. (Bu4N)2[Fe4S4(LS3)(CF3SO3)]. To a solution of 214 mg (0.116 mmol) of (Bu4N)2[Fe4S4(LS3)(SEt)]41 in 15 mL of acetonitrile was added a solution of 28.7 mg (0.115 mmol) of (Et3NH)(CF3SO3) in 15 mL of acetonitrile. The reaction mixture was stirred for 10 min and filtered. Slow removal of solvent afforded the pure product as a black crystalline solid in quantitative yield. 1H NMR (CD3CN): δ 8.24 (5-H), 7.08 (2′-H), 6.81 (3′-H), 5.02 (2-H), 3.93 (4-Me), 3.89 (6-Me), 2.22 (4′Me). (Bu4N)2[Fe4Se4(LS3)(SEt)]. To a solution of 1.00 g (0.788 mmol) of (Bu4N)2[Fe4Se4(SEt)4]42 in 150 mL of acetonitrile was added 786 (34) (a) Berg, J. M.; Holm, R. H. In Iron-Sulfur Proteins; Spiro, T. G., Ed.; Wiley: New York, 1982; Chapter 1. (b) Holm, R. H.; Ciurli, S.; Weigel, J. A. Prog. Inorg. Chem. 1990, 29, 1. (35) Hagen, K. S.; Watson, A. D.; Holm, R. H. J. Am. Chem. Soc. 1983, 105, 3905. (36) Srivastava, K. K. P.; Surerus, K. K.; Conover, R. C.; Johnson, M. K.; Park, J.-B.; Adams, M. W. W.; Mu¨nck, E. Inorg. Chem. 1993, 32, 927. (37) Finnegan, M. G.; Conover, R. G.; Park, J.-B.; Zhou, Z. H.; Adams, M. W. W.; Johnson, M. K. Inorg. Chem. 1995, 34, 5358. (38) (a) Ciurli, S.; Ross, P. K.; Scott, M. J.; Yu, S.-B.; Holm, R. H. J. Am. Chem. Soc. 1992, 115, 5415. (b) Zhou, J.; Scott, M. J.; Hu, Z.; Peng, G.; Mu¨nck, E.; Holm, R. H. J. Am. Chem. Soc. 1992, 114, 10843. (39) Zhou, J.; Holm, R. H. J. Am. Chem. Soc. 1995, 117, 11353. (40) Surerus, K. K.; Kennedy, M. C.; Beinert, H.; Mu¨nck, E. Proc. Natl. Acad. Sci. U.S.A. 1989, 86, 9846. (41) (a) Stack, T. D. P.; Holm, R. H. J. Am. Chem. Soc. 1988, 110, 2484. (b) Liu, H. Y.; Scharbert, B.; Holm, R. H. J. Am. Chem. Soc. 1991, 113, 9529. (42) Yu, S.-B.; Papaefthymiou, G. C.; Holm, R. H. Inorg. Chem. 1991, 30, 3476.

The Cuboidal Fe3S4 Cluster mg (0.828 mmol) of L(SH)3.43 The reaction mixture was stirred for 12 h, during which it was subjected to dynamic vacuum for 10 min every ca. 4 h to remove ethanethiol. The solvent was removed in vacuo; the microcrystalline solid residue was dissolved in 50 mL of acetonitrile and the solution was filtered. Volume reduction of the filtrate caused separation of product as 1.52 g (95%) of black microcrystals. 1H NMR (CD3CN): δ 15.57 (SCH2), 8.41 (5-H), 7.02 (2′-H), 6.66 (3′-H), 5.55 (2-H), 4.20 (4-Me), 3.93 (6-Me), 2.21 (4′-Me). (Bu4N)2[Fe4Se4(LS3)(CF3SO3)]. To a solution of 220 mg (0.108 mmol) of (Bu4N)2[Fe4Se4(LS3)(SEt)] in 50 mL of acetonitrile was added a solution of 28.6 mg (0.113 mmol) of (Et3NH)(CF3SO3) in 5 mL of acetonitrile. The reaction mixture was stirred for 10 min and filtered. Slow removal of solvent afforded the pure product as a black crystalline solid in quantitative yield. 1H NMR (CD3CN): δ 8.51 (5-H), 6.97 (2′-H), 6.62 (3′-H), 5.49 (2-H), 4.38 (4-Me), 4.04 (6-Me), 2.20 (4′Me). (Et4N)3[Fe4S4(LS3)(Meida)]. To a solution of 195 mg (0.101 mmol) of (Bu4N)2[Fe4S4(LS3)(CF3SO3)] in 10 mL of acetonitrile was added 43 mg (0.106 mmol) of (Et4N)2(Meida) in 5 mL of acetonitrile. The reaction mixture was stirred for 2 h and filtered. Removal of solvent in vacuo gave a black microcrystalline solid, which was collected and washed with 10 mL of THF and 10 mL of acetone to afford 176 mg (95%) of pure product. Absorption spectrum (Me2SO): λmax (M) 365 (sh, 38 300), 472 (21 200) nm. 1H NMR (Me2SO): δ 8.55 (5-H), 7.09 (2′-H), 6.58 (3′-H), 4.47 (6-Me), 3.89 (4-Me), 3.49 (CH2), 3.24 (NMe), 2.19 (4′-Me), 1.65 (CH2). (Bu4N)3[Fe4S4(LS3)(citrate)]. To a solution of 150 mg (0.078 mmol) of (Bu4N)2[Fe4S4(LS3)(CF3SO3)] in 5 mL of acetonitrile was added a solution of 45 mg (0.078 mmol) of (Et4N)3(citrate) in 5 mL of acetonitrile. The mixture was stirred for 1 h, over which time a black microcrystalline solid separated. This material was collected by filtration and washed with THF and acetone, giving 100 mg (65%) of product. 1H NMR (Me2SO): δ 8.55 (5-H), 7.14 (2′-H), 6.57 (3′-H), 4.47 (6-Me), 3.89 (4-Me), 3.19 (sh, CH2), 2.14 (4′-Me), 1.37 (CH2), 0.83 (CH2). (Et4N)3[Fe3S4(LS3)]. To a solution of 210 mg (0.108 mmol) of (Bu4N)2[Fe4S4(LS3)(CF3SO3)] in 20 mL of acetonitrile was added a solution of 176 mg (0.434 mmol) of (Et4N)2(Meida) in 5 mL of acetonitrile. The reaction was stirred for 30 min, during which a black microcrystalline solid separated. This material was collected by filtration and washed with 10 mL of THF, 10 mL of acetone, and 5 mL of acetonitrile to yield 140 mg (80%) of pure product. Absorption spectrum (Me2SO): λmax (M) 365 (sh, 28 300), 475 (br sh, 13 100) nm. 1H NMR spectrum (Me2SO): δ 15.48 (5-H), 11.66 (6-Me), 8.51 (4-Me), 7.06 (2′-H), 6.38 (3′-H), 2.36 (4′-Me). Anal. Calcd for C75H105Fe3N3S13: C, 55.02; H, 6.46; Fe, 10.23; N, 2.57; S, 25.46. Found: C, 54.86; H, 6.74; Fe, 10.35; N, 2.53; S, 25.38. (Et4N)3[Fe3Se4(LS3)]. To a solution of 200 mg (0.094 mmol) of (Bu4N)2[Fe4Se4(LS3)(CF3SO3)] in 10 mL of acetonitrile was added a solution of 153 mg (0.377 mmol) of (Et4N)2(Meida) in 5 mL of acetonitrile. The solution was stirred for 10 min, filtered, and allowed to stand overnight. The black crystalline solid was collected by filtration and washed with THF and acetone to give 150 mg (87%) of product. Absorption spectrum (Me2SO): λmax (M) 357 (sh, 77 000), 465 (br sh, 35 200). 1H NMR (Me2SO, 297 K): δ 15.94 (5-H), 12.12 (6-Me), 9.97 (4-Me), 7.01 (2′-H), 6.25 (3′-H), 2.24 (4′-Me). Anal. Calcd for C75H105Fe3N3S9Se4: C, 49.48; H, 5.81; Fe, 9.20; N, 2.30; S, 15.85; Se, 17.35. Found: C, 49.29; H, 5.99; Fe, 9.15; N, 2.25; S, 16.03; Se, 17.21. (Et4N)2[Fe(Meida)2]. This compound was generated in solution by the reaction of 1 equiv of Fe(CF3SO3)2 with 2 or more equiv of (Et4N)2(Meida); 1H NMR spectra were recorded for purpose of identifying [Fe(Meida)2]2- formed in the iron abstraction reactions of Figure 2. CD3CN: δ 45.2 + 44.1 (2, Me + CHb), 70.9 (1, CHa). DMF/CD3CN (1:1 v/v): δ 44.0 (2), 72.4 (1). (CD3)2SO: δ 45.5 + 44.6 (2), 70.9 (1). X-ray Structure Determination. Hexagonal red-black plates of (Et4N)3[Fe3S4(LS3)] were grown by vapor diffusion of THF into a DMF solution of the cluster salt. A suitable single crystal was coated with grease and attached to a glass fiber. Diffraction data were collected at -50 °C using a Siemens SMART area diffractometer at Northern Illinois University. Lattice parameters were obtained from least-squares (43) Stack, T. D. P.; Weigel, J. A.; Holm, R. H. Inorg. Chem. 1990, 29, 3745.

J. Am. Chem. Soc., Vol. 118, No. 8, 1996 1969

Figure 2. Scheme for the synthesis of Fe3Q4 cuboidal clusters 11 and 12 (Q ) S, Se); intermediate clusters 6-10 have been isolated. The structures of 8-10 are consistent with NMR spectra and considered probable in solution. (3LS ) LS3). Table 1. Crystallographic Data for (Et4N)3[Fe3S4(LS3)]‚MeCN formula formula wt crystal system space group Z a, Å b, Å c, Å V, Å3 Fcalc, g/cm3 T, K R,a wR2b a

C77H108Fe3N4S13 1674.13 orthorhombic P212121 4 14.8042(2) 18.3989(3) 33.1771(2) 9036.8(2) 1.328 223 0.0793, 0.1771

R ) ∑||Fo| - |Fc||/∑|Fo|. b wR2 ) {∑[w(Fo2 - Fc2)2]/∑[w(Fo2)2]}1/2

analysis of 100 machine-centered reflections with 10° e 2θ e 20°. The collected image files were converted to a raw intensity data file using the Siemens SAINT program, and then converted to structure factor amplitudes and their esd’s by correction for scan speed, background, and Lorentz and polarization effects using SHELXTL PLUS. The space group was identified by systematic absences. Data collection and crystal parameters are listed in Table 1. The structure was solved by direct methods and was refined by standard least-squares and Fourier techniques. Hydrogen atoms were assigned ideal locations and given the uniform value Biso ) 0.8 Å2. The asymmetric unit consists of three cations, one anion, and one acetonitrile solvate molecule. In the last cycle of refinement, all parameters shifted by 90% in

[Fe4S4(LS3)(SEt)]2- + 2(Meida)2- f [Fe3S4(LS3)]3- + [Fe(Meida)2]2- + EtS- (3) situ conversion to 11 with 4 equiv of abstracting ligand; several unidentified signals are observed in the 6-7 ppm region but there is no indication of the formation of another paramagnetic cluster type. On a preparative basis, triflate cluster 6 leads to a purer product, and thus is the preferred precursor to the cuboidal cluster. (c) Stability. We have previously shown that the reaction of [Fe4S4(LS3)(S-t-Bu)]2- with >4 equiv of t-BuSNa in Me2SO resulted in decoordination of the LS3 ligand and formation of [Fe4S4(StBu)4]2- in high yield.41a In this work, a similar reaction 4 of 4 with NaSEt afforded quantitative formation of the cubane product. The analogous reaction 5 proceeds,

[Fe4S4(LS3)(SEt)]2- + 3NaSEt f [Fe4S4(SEt)4]2- + Na3(LS3) (4) [Fe3S4(LS3)]3- + 3NaSEt f [Fe3S4(SEt)3]3- + Na3(LS3) (5) although not stoichiometrically, and provides a means of studying the stability of the product cuboidal clusters. A solution of 11 (6.1 µmol in 0.5 mL) was treated with a solution of NaSEt (30 µmol in 0.1 mL). A spectrum taken at 10 min reaction time showed the relative percentages of the cluster products to be 65% [Fe3S4(SEt)3]3- and 35% [Fe4S4(SEt)4]2-; no starting cluster remained. After 1 h, these amounts decreased and increased by 10%, respectively, and a trace amount of [Fe2S2(SEt)4]2- appeared. At 1-50 h, the direction of these changes continued and a small quantity of linear [Fe3S4(SEt)4]3appeared and decayed, until at 50 h there remained 11% [Fe3S4(SEt)3]3-, 35% [Fe2S2(SEt)4]2-, and 54% [Fe4S4(SEt)4]2-. The means of formation of the non-cuboidal clusters is obscure. What is evident, however, is that [Fe3S4(SEt)3]3-, in the presence of bi-, tri-, and tetranuclear clusters which may be derived from it, is moderately unstable and decays to known clusters. This cluster appears sufficiently stable to be isolable, but under conditions requiring separation from other clusters. We have not observed formation of [Fe3S4(SEt)3]3-, by reactions 3 or 5 or any other means, without accompanying formation of other cluster(s). Cluster 11 is stable for at least 1 week, and 12 for 2-3 days, in anaerobic solutions at ambient temperature. Thus, the auspicious effect of the LS3 ligand system in promoting both the formation and stability of the [Fe3S4]0 core is demonstrated. (62) Because of large line widths of most resonances, percentages of species obtained by integration in this and related experiments are semiquantitative only. (63) Carney, M. J.; Papaefthymiou, G. C.; Spartalian, K.; Frankel, R. B.; Holm, R. H. J. Am. Chem. Soc. 1988, 110, 6084 and references therein.

Figure 8. Structure of [Fe3S4(LS3)]3- as its Et4N+ salt, showing the atom numbering scheme for Fe and S atoms and for one ligand arm and leg, and 50% probability ellipsoids. Lower: Perspective view of the entire structure roughly perpendicular to the idealized C3 axis, which contains S(1) and bisects the central benzene ring carrying S(13,23,33). Upper: View along the idealized C3 axis emphasizing the ∆ absolute configuration.43

Cluster Reconstitution. As a further structure proof in solution, we have demonstrated that reaction 6 proceeds

[Fe3Q4(LS3)]3- + [FeCl4]2- f [Fe4Q4(LS3)Cl]2- + 3Cl- (6) quantitatively in Me2SO to yield the chloride clusters with the following chemical shifts: 14, 3.79 (4-Me), 3.90 (6-Me), 8.22 (5-H); 15, 4.28 (4-Me), 4.04 (6-Me), 8.48 (5-H). The values are identical with those of the authentic clusters41a,43 in the same solvent. The minimal reaction is the reverse of preparative reaction 1, Viz. [Fe3Q4]0 + Fe2+ f [Fe4Q4]2+. Solid State Structure of [Fe3S4(LS3)]3-. The compound (Et4N)3[11]‚MeCN crystallizes in an orthorhombic space group with no imposed symmetry. Two views of the structure of the entire anion are available in Figure 8 and the core structure is shown in more detail in Figure 9. It is immediately evident

1974 J. Am. Chem. Soc., Vol. 118, No. 8, 1996

Zhou et al. Table 3. Conformational Analysis of [Fe3S4(LS3)]3- (deg)a conformation

ring

ψ (ring 0/ring n)

θ (tilt)

φ (cant)

ababab

1 2 3 4 5 6

66.63 69.62 70.42 106.76 105.53 97.59

51.88 44.28 52.96 -85.19 -64.29 -71.10

27.74 44.04 30.87 -46.91 -20.59 -31.96

a Tilt and cant angles are defined by the dihedral angles formed by the shaded atoms relative to a planar reference conformation with θ ) φ ) 0°.

Figure 9. Structure of the [Fe3S4]0 core of [Fe3S4(LS3)]3-, with 50% probability ellipsoids and the atom labeling scheme. Table 2. Selected Interatomic Distances (Å) and Angles (deg) for [Fe3S4(LS3)]3Fe(1)-S(1) Fe(2)-S(1) Fe(3)-S(1) mean of 3

2.310(2) 2.273(3) 2.333(2) 2.31(3)

Fe(1)-Fe(2) Fe(1)-Fe(3) Fe(2)-Fe(3) mean of 3

2.712(2) 2.665(2) 2.731(2) 2.70(3)

Fe(1)-S(2) Fe(1)-S(3) Fe(2)-S(3) Fe(2)-S(4) Fe(3)-S(4) Fe(3)-S(2) mean of 6

2.274(2) 2.251(3) 2.242(2) 2.265(3) 2.250(3) 2.275(3) 2.26(2)

Fe(1)-S(11) Fe(2)-S(21) Fe(3)-S(31) mean of 3

2.316(3) 2.310(3) 2.327(3) 2.318(9)

S(11)-C(111) S(21)-C(211) S(31)-C(311) mean of 3

1.746(8) 1.774(9) 1.746(10) 1.76(2)

Fe(2)-S(1)-Fe(1) Fe(2)-S(1)-Fe(3) Fe(1)-S(1)-Fe(3)

72.56(8) 72.70(8) 70.05(7)

S(2)-Fe(1)-S(3) S(3)-Fe(2)-S(4) S(4)-Fe(3)-S(2)

112.5(1) 111.5(1) 113.8(1)

S(2)-Fe(1)-S(1) S(3)-Fe(1)-S(1) S(3)-Fe(2)-S(1) S(4)-Fe(2)-S(1) S(4)-Fe(3)-S(1) S(2)-Fe(3)-S(1)

106.06(9) 102.17(9) 103.59(10) 103.02(9) 101.63(9) 105.27(9)

Fe(3)-Fe(1)-Fe(2) Fe(1)-Fe(3)-Fe(2) Fe(1)-Fe(2)-Fe(3)

61.03(4) 60.34(4) 58.63(4)

Fe(1)-S(3)-Fe(2) Fe(2)-S(4)-Fe(3) Fe(3)-S(2)-Fe(1)

74.27(8) 74.42(8) 71.71(8)

that the desired cuboidal cluster has been achieved. The structures of only two other LS3 clusters have been determined, (Ph4P)2[14]41a and (Ph4P)2[15]‚2DMF;43 both have a common chloride ligand at the unique site. The latter crystallizes in trigonal space group P3 with three one-third anions in the asymmetric unit. We shall have occasion to compare the present structure with these. (a) Conformation. We have developed a conformational analysis for substituted benzenes whose phenyl substituents are connected to the central ring (ring 0) by one atom.41a The molecular conformation of 11 is describable at two levels. First, the cluster is found in the ababab conformation, in which alternate phenylthio substitutents are above and below the plane of ring 0. The three ligand arms with rings 1-3 coordinate the Fe3S4 core with atoms S(11,21,31), and are buttressed into position by the three p-tolylthio legs. The six connecting sulfur atoms of ring 0 are below it (away from the core) at a mean distance of 0.19 Å. This displacement contributes to the placement of S(1) at 3.77 Å above the center of ring 0, compared to 3.74 Å for 14. The van der Waals contact distance between ring 0 and S(1) is estimated to be 3.44 Å. The core substantially occupies the ligand cavity whose walls are defined by the inward edges of rings 1-3. At a second level, the conformation is quantitatively described by the dihedral angles in Table 3. the inequality within each set of the angles ψ between ring 0 and

rings 1-3 (arms) and rings 4-6 (legs) is the simplest indication that the ligand conformation lacks C3 symmetry. The conformation is precisely described by the angles θ (tilt) and φ (cant), which also indicate departure from trigonal symmetry and convey the orientation of rings 1-6 with respect to ring 0. The binding of the cuboidal core to the ligand renders the cluster chiral. However, the sign of neither the tilt nor cant angle necessarily correlates with absolute configuration. Given the criteria developed previously,43 the absolute configuration of the cluster in the crystal of (Et4N)3[11]‚MeCN examined is ∆. A clockwise trigonal twist about the idealized C3 axis viewed from the S(2-4) core face recovers the Λ enantiomer. Earlier, cluster 15 was found in an inversion-twinned crystal with the ratio ∆:Λ ) 2:1.43 (b) Core. Under idealized C3V symmetry, the bond distances and angles of a cuboidal Fe3S4 divide into 3 + 6Fe-S, 3FeFe, (3 + 3)Fe-S-Fe, (3 + 6)S-Fe-S, and 3Fe-Fe-Fe. Metric data are arranged in this manner in Table 2. The data reveal departures from this symmetry in a manner suggestive of a pseudomirror plane containing atoms Fe(2), S(1), and S(2). Bond distances and angles are unexceptional for Fe-S clusters. The most interesting structural aspect is the relationship between the cubane and cuboidal cores, where the issue is the extent of change when an iron atom is in actuality removed from the former to generate the latter. For this purpose, we take 14 as the reference. Mean values of independent structural parameters for the [Fe3S4]0 core of 11 and the [Fe4S4]2+ core of 14 are compared in Figure 10. The latter lacks the elongated tetragonal distortion common to such cores34a,63 and instead approaches Td symmetry, under which its metric data have been averaged. The absent iron atom in the depiction is understood to be that at the unique site. The comparison, made in terms of mean values of angles and distances, carries the caveat that the mean iron oxidation states of the cores differ (Fe2.67+ in 11, Fe2.50+ in 14). Given this difference, we are unable to explain why the terminal Fe-S distances are longer in 11 (2.32 Å) than in 14 (2.26 Å). With almost no exceptions, bond lengths decrease as the oxidation state increases in the series [Fe4S4]1+,2+,3+,34a,64 i.e., as the Fe3+ character of the core increases.65 Removal of an iron atom from a cubane core might be expected to cause relaxation of the structure to a more open, or splayed, configuration. In this case, the remaining µ3-S atom would experience increased Fe-S(1)-Fe bond angles and the three S-Fe-S angles would expand. Evidence for this effect (64) (a) O’Sullivan, T.; Millar, M. M. J. Am. Chem. Soc. 1985, 107, 4096. (b) Carney, M. J.; Papaefthymiou, G. C.; Frankel, R. B.; Holm, R. H. Inorg. Chem. 1989, 28, 1497 and references therein.

The Cuboidal Fe3S4 Cluster

Figure 10. Metric features of the core structures of [Fe4S4(LS3)Cl]2- 41a (mean bond distances and angles averaged under idealized Td symmetry; one Fe atom removed from core), [Fe3S4(LS3)]3- (mean bond distances and angles averaged under idealized C3V symmetry), and D. gigas Fd II.5b The small numbers correspond to the atom labeling scheme of Figure 9 and of the protein structure. Certain angles are shown outside the protein cluster, which is depicted after Kissinger et al.5

is found in the S-Fe-S angle of 112.6° vs 103.6° in the cubane core. The angles at µ3-S have contracted by a small amount (2.6°) in passing to the cuboidal form. Overall, the cubane and cuboidal cores are closely related metrically. Other than the 9.0° angle increase noted, the principal dimensional difference in the two cores is the 0.07-Å decrease in Fe-Fe separations in forming the cuboidal species. The average values of all Fe-S distances in the two cores are indistinguishable (2.28, 2.29 Å). These bond lengths are symmetry-differentiated in the cuboidal core where, as expected, Fe-(µ2-S) bonds are shorter than Fe(µ3-S) bonds (by 0.05 Å). In the structure of oxidized AV Fd I, the reported average values Fe-Fe ) 2.68 Å and Fe-(µ3-S)-Fe ) 70.5° 6a are close to the corresponding values in 11 (2.70 Å, 71.8°). The structure of this protein has also been determined in the oxidized and dithionite-reduced ([Fe3S4]0) forms at pH 8 and 6.6c The cluster structure is the same in the four structures, with some evidence adduced for protonation of the reduced form at acid pH. Explicit dimensional data for reduced clusters were not reported. Included in Figure 10 are the distance and angle data for the [Fe3S4]1+ core of Dg Fd II.5b These data were not averaged because of the large spread in certain parameters that would be identical under trigonal symmetry. Despite core distortions and a different oxidation state, it is clearly evident that structures of the synthetic and protein-bound cores are substantially the same. Kissinger et al.5 have emphasized the close dimensional relationship between the Dg Fd II cluster and those of typical (65) It has not escaped our attention that the pattern of Fe-Fe separations in 11 (Table 2) is such as to approximate an isosceles triangle. On the basis of this pseudosymmetry, Fe(1,3) would be the delocalized pair and Fe(2) the Fe3+ site. However, the Fe-Fe distances themselves are not necessarily consistent with this picture. Further, the terminal Fe-S bond lengths, which are generally sensitive to the oxidation states of the Fe atom in Fe-S clusters as noted above, cover a range of j0.02 Å, and thus do not reflect differences in oxidation states. Additional structures of [Fe3S4]0 clusters are being sought to examine the relationship between metric features and electronic structure.

J. Am. Chem. Soc., Vol. 118, No. 8, 1996 1975

Figure 11. Cyclic voltammograms (50 mV/s) of [Fe4Q4(LS3)(Meida)]3in acetonitrile with Q ) S (upper) and Se (lower). Peak potentials are indicated; x ) slight impurity of [Fe4S4(LS3)(OSO2CF3)]2-.

Fd proteins in the [Fe4S4]2+ state. Stout6a has concluded that “there is no distinguishable difference” between the [Fe3S4]1+ and [Fe4S4]2+ clusters of AV Fd I except for the number of atoms. The lack of a major structural change contributes to the facility of removal of Fe2+ from the cubane core and the ready reconstitution of the core in reaction 6. The same point applies to the binding of extrinsic metal ions to the synthetic39 and protein-bound7,10,36,37 cores to afford the heterometal cubanetype MFe3S4 clusters. Electron Transfer Reactions. Relevant electron transfer series and redox potentials are collected in Table 4. Unless noted otherwise, the processes are chemically reversible (ipa/ipc ≈ 1) one-electron reactions. Shown in Figure 11 are cyclic voltammograms of the two immediate precursors to the cuboidal clusters, the cubanes 8 and 9, which exhibit the three-membered series 7. The less negative potential for reduction of [Fe4Se4]2+ clusters is precedented.42,59a Amongst LS3 species this property is further illustrated by the values for 14 and 15 in DMF, which differ by 80 mV. The more negative potentials for 8 and 9 are most likely due to their 3- charge. The conversions 8 f 11 and 9 f 12 afford cuboidal clusters that manifest the chemically reversible, three-membered electron transfer series 8; these processes approach (at 50 mV/s) the criterion ∆Ep ) 59 mV for an electrochemically reversible one-electron reaction. Although of equal overall charge, the potentials of 11/12, shown in the voltammograms of Figure 12, are considerably more negative than those of 8/9, an effect we ascribe to the differences in core charges in the two sets of clusters. For example, on a charge basis it should be more difficult to reduce a [Fe3S4]0 than a [Fe4S4]2+ core at near-parity of terminal ligation. Inasmuch as the [Fe3S4]1+,0 states are known in proteins and [Fe3S4]0 has now been synthesized, the most interesting result is the generation of the [Fe3S4]1- state, detected for the first time in the absence of a stabilizing cation as in certain heterometal cubanes 3. The decidedly negative potential for the formation of this oxidation state is suggestive of the need for cluster protonation in stabilization of the protein-bound

1976 J. Am. Chem. Soc., Vol. 118, No. 8, 1996

Zhou et al.

Table 4. Cluster Electron Transfer Series and Redox Potentials E1/2, Va cluster 3-

[Fe4S4(LS3)(Meida)] (8) [Fe4Se4(LS3)(Meida)]3- (9) [Fe4S4(LS3)Cl]2- (14) [Fe4Se4(LS3)Cl]2- (15) [Fe3S4(LS3)]3- (11) [Fe3Se4(LS3)]3- (12) [Fe4S4(LS3)(t-BuNC)]1- (16) [(OC)3MoFe3S4(LS3)]3- (17) [(OC)3MoFe3S4(Smes)3]3- (18) [(OC)3MoFe3S4(SEt)3]3- (19)d a

solvent

series

MeCN MeCN DMF CH2Cl2b DMF MeCN MeCN CH2Cl2b MeCN MeCN CH2Cl2

(7a) (7b)

0/1-

1-/2-

2-/3-

3-/4-

-0.21 -0.27

-1.25 -1.20 -1.03 -1.08 -0.95 -0.79 -0.80

-1.72 -1.67

∼-0.20c -0.31 -0.40

-0.78 -0.86 -0.94

+0.12 (8a) (8b) (9) (10a) (10b) (10c)

-0.18

-1.08

298 K, vs SCE. E1/2 ) (Epc + Epa)/2. b Reference 50. c Poorly defined feature. d For additional data on clusters of this type, cf. ref 51.

bound [Fe3Q4]0 clusters.2,3,13,26,28,40,66-68 Mo¨ssbauer spectra recorded in the absence of an applied magnetic field consist of two quadrupole doublets with intensity ratio 2:1. The less intense doublet typically has an isomer shift, δ ) 0.30-0.35 mm/s, slightly larger than those observed for a ferric site with tetrahedral sulfur coordination, while the more intense doublet has a shift, δ ) 0.46-0.49 mm/s, suggestive of a valencedelocalized Fe2+Fe3+ pair. In the presence of applied magnetic fields, the low-temperature Mo¨ssbauer spectra display intricate and well-resolved magnetic patterns whose features reflect the properties of an S ) 2 spin system with zero-field splitting parameters D ≈ -2.5 cm-1 and E/D ) 0.20-0.25. The spectra have been analyzed with the spin Hamiltonian 7 (for details, see ref 13), where the quadrupole interactions are given by eq 8. The electron spin S is the cluster spin, D and E are the axial

[

]

E 1 H ) D Sz2 - S(S + 1) + (Sx2 - Sy2) + βH‚g‚S + 3 D 3

[S‚A(i)‚I(i) - gnβnH‚I(i) + HQ(i)] ∑ i)1 HQ(i) )

Figure 12. Cyclic voltammograms (50 mV/s) of [Fe3Q4(LS3)]3- in acetonitrile with Q ) S (upper) and Se (lower). Peak potentials are indicated; x ) impurity.

[Fe3S4]1-,2- states at accessible potentials in water (Figure 1). Electron transfer series 9 and 10 based on spin-isolated clusters 16-19 cover at least the [Fe3S4]0,1- core oxidation states that are traversed in series 8a. The site of oxidation is perhaps ambiguous and could be either the cuboidal fragment or lowspin Fe(II) (16) or Mo(O) (17-19). However, reduction potentials larger than ca. -1.0 V are not reasonably assignable to such sites. At parity of overall charge in series 8a and 10, stabilization of the [Fe3S4]1- state is quite evident. The most exact comparison is with 11 and 17, for which the stabilization is 0.94 V in acetonitrile. We find this value unexpectedly large, given that in the analogous clusters 1839 and 19,51 X-ray structural results clearly show that the Mo(CO)3 group is weakly bonded to the [Fe3S4]0 fragment. Series 10b closely approaches electrochemical reversibility; its most reduced member is a potential source of the [Fe3S4]1- core without LS3 stabilization by displacement of Mo(CO)3 and trapping with metal ions. Mo1 ssbauer Spectroscopy and Electronic Structure. Before presenting data pertinent to electronic structure, we summarize briefly the salient features of the Mo¨ssbauer spectra of protein-

eQVζζ(i) 2 {3Iζ(i) - I(I + 1) + η(i)[I2ξ(i) - I2η(i)]} 12

(7)

(8)

and rhombic zero-field splitting parameters, respectively, and g is the electronic g tensor. The magnetic hyperfine interactions of the three sites of an [Fe3S4]0 cluster are described by the tensors A(i), where i ) 1, 2, 3 sums over the three iron sites. We have found that the ζ-axes of the electric field gradient tensors (principal axes components Vξξ, Vηη, Vζζ) of the individual sites are rotated relative to the z-axis of the zerofield splitting tensor, and we describe this rotation by the polar angle β. At 4.2 K, the electronic spins of all the Fe3S4 clusters studied to date relax slowly on the time scale of Mo¨ssbauer spectroscopy (≈10 MHz). Under these conditions, the salient features of the low-temperature spectra are determined by the expectation value of the spin of the lowest electronic level. A plot of 〈Si〉 along the principal axes of the ZFS tensor shown in Figure 13 suggests the strategy to be used for the analysis of the spectra. Thus, at low field 〈Sz〉 saturates readily at 〈Sz〉 ) -2, while 〈Sx〉 and 〈Sy〉 remain small. Consequently, the lowfield (H < 2.0 T) spectra of each iron site are dominated by the magnetic hyperfine field along z, Hint(i) ) -〈Sz〉Az(i)/gnβn, and by the component of the electric field gradient tensor along (66) Surerus, K. K.; Chen, M.; van der Zwaan, W.; Rusnak, F. M.; Kolk, M.; Duin, E. C.; Albracht, S. P. J.; Mu¨nck, E. Biochemistry 1994, 33, 4980. (67) Hu, Z.; Jollie, D.; Burgess, B. K.; Stephens, P. E.; Mu¨nck, E. Biochemistry 1994, 33, 14475. (68) Teixeira, M.; Moura, I.; Xavier, A. V.; Moura, J. J. G.; LeGall, J.; DerVartanian, D. V.; Peck, H. D., Jr.; Huynh, B. H. J. Biol. Chem. 1989, 264, 16435.

The Cuboidal Fe3S4 Cluster

J. Am. Chem. Soc., Vol. 118, No. 8, 1996 1977

Figure 13. Plot of 〈Sx〉, 〈Sy〉, and 〈Sz〉 for the lowest electronic level of the S ) 2 system for D ) -2.5 cm-1, E/D ) 0.25, and gx ) gy ) gz ) 2.0. 〈Sx〉 is the expectation value of S with the magnetic field applied along the x-direction. The jump in 〈Sy〉 reflects a level crossing.

Hint. As the applied field is increased, the x and y components of the A tensors become important. By following the increase of the magnetic hyperfine field with increasing applied field, the ZFS parameters D and E/D can be determined. Thus, by a judicious choice of experimental conditions, a substantial number of fine structure and hyperfine structure parameters can be determined with good precision. Finally, since the g-values of eq 7 are close to g ) 2, the Mo¨ssbauer spectra are quite insensitive to g. Figures 14D and 14B show 4.2 K zero-field Mo¨ssbauer spectra of clusters 11 and 12, respectively. For comparison, the corresponding spectrum of Dg Fd II is shown in Figure 14C. The spectra of the synthetic clusters exhibit intensity patterns essentially identical with those observed for proteins containing the [Fe3S4]0 26-28 and [Fe3Se4]0 40 clusters. By using a Fourier transform deconvolution69 that removes the line width contribution of the 57Co source, one can frequently improve the resolution of the Mo¨ssbauer spectra. As can be seen from the spectrum of Figure 14A, application of this technique to the spectrum of Figure 14B shows that the quadrupole splittings and isomer shifts of the sites P2 and P3 (the delocalized Fe2+Fe3+ pair) differ slightly. The solid lines in Figures 14B and 14D are least-squares fits that yielded the parameters for ∆EQ and δ listed in Table 5.70 As outlined for the analysis of Dg Fd II,13 the ground state of the [Fe3S4]0 cluster has an electron configuration consisting of a valence-delocalized Fe2+Fe3+ pair and a trapped-valence Fe3+ site. The Mo¨ssbauer study of Dg Fd II has also revealed an excited configuration, X, that becomes populated at temperatures above 25 K. Configuration X, represented by one quadrupole doublet with ∆EQ ≈ 0.9 mm/s and δ ) 0.45 mm/s, seems to reflect a situation where the excess electron is evenly delocalized among the three iron sites.13 Configuration X has not been reported for other [Fe3S4]0-containing proteins. In order to assess whether such an excited state becomes thermally accessible for the two synthetic clusters, we have studied their (69) Huynh, B. H.; Mu¨nck, E.; Orme-Johnson, W. H. Biochim. Biophys. Acta 1979, 527, 192. (70) Analysis of the spectra of all data sets suggests that 11 and 12 contain a ca. 5% contaminant of [Fe4S4]2+ and [Fe4Se4]2+ species, respectively. Both the zero-field and high-field spectra indicate additional absorbance in the spectral range where (diamagnetic) [Fe4Q4]2+ clusters typically absorb. For instance, a fit with free intensities to the spectrum of Figure 14B yields 31% and 69% for the intensities of site 1 and the sum of sites P2 and P3, respectively, rather than the expected 33.3% and 66.7%. While these differences could be attributed to different recoiless fractions of the sites, this interpretation would not explain the additional absorption in the “diamagnetic regions” of the high-field spectra.

Figure 14. Mo¨ssbauer spectra of polycrystalline (Et4N)3[Fe3S4(LS3)] and (Et4N3)[Fe3Se4(LS3)] and of Dg Fd II recorded in zero applied magnetic field. (B) 4.2 K spectrum of (Et4N)3[Fe3Se4(LS3)]. The spectrum in (A) was obtained from the data in (B) by removing the line width contribution of the 57Co source. (C) 4.2 K spectrum of Dg Fd II; the solid line indicates the contribution of the delocalized Fe2+Fe3+ pair, and the innermost doublet belongs to the Fe3+ site. (D and E) Spectra of (Et4N)3[Fe3S4(LS3)]. The solid lines in (B), (D), and (E) are least-squares fits to the data assuming three doublets. Typical line width parameters were 0.28-0.32 mm/s (full width at half maximum). The spectra of the synthetic clusters each contain ≈5% contaminant of a [Fe4Q4]2+ species; this contribution (∆EQ ) 1.2 mm/s, δ ) 0.45 mm/s) has been subtracted from the raw data. Table 5. Quadrupole Splittings and Isomer Shifts of [Fe3Q4(LS3)]3- (mm/s) [Fe3S4]0

[Fe3Se4]0

T (K)

site

∆EQ

δ

∆EQ

δ

4.2

P1 P2 Fe3+ P1 P2 Fe3+ P1 P2 Fe3+

1.48 1.18 0.51 1.45 1.11 0.49 1.38 1.09 0.52

0.49 0.48 0.35 0.45 0.44 0.32 0.43 0.42 0.30

1.45 1.20 0.47 1.26 0.94 0.45 1.24 0.91 0.45

0.52 0.49 0.39 0.48 0.46 0.39 0.46 0.45 0.37

150 200

zero-field Mo¨ssbauer spectra at higher temperatures. The 200 K spectrum of 11, shown in Figure 14E, exhibits the same 2:1 intensity pattern as the 4.2 K spectrum, showing that configuration X is not measurably populated at 200 K. The same result was obtained for 12. The temperature dependencies of ∆EQ and δ are listed in Table 5. We have recorded Mo¨ssbauer spectra at 4.2 K in applied fields up to 8.0 T. Representative spectra are shown in Figures 15 and 16. Both complexes were observed to exhibit resolved magnetic hyperfine interaction in fields as low at 0.1 T, showing that the electronic spin fluctuates slowly on the time scale of Mo¨ssbauer spectroscopy. (The two lowest levels of the S ) 2 system are separated by 0.3 cm-1, and their spin expectation values are equal but have opposite signs. In the fast fluctuation

1978 J. Am. Chem. Soc., Vol. 118, No. 8, 1996

Figure 15. Mo¨ssbauer spectra of (Et4N)3[Fe3S4(LS3)] (A, C, D) recorded in magnetic fields applied parallel to the observed γ-radiation. For comparison, the 6.0 T spectrum of Dg Fd II is shown (B). Solid lines are theoretical spectra generated from eq 7 using the parameters in Table 6 for the case βA ) 0; P1 and P2 are components of the delocalized pair and Fe3+ is the ferric site.

limit only quadrupole doublets would be observed for H ) 0.1 T.) It can be seen that the applied field spectra of both synthetic complexes are remarkably similar to those observed for proteinbound [Fe3S4]0 (Figure 15B) and [Fe3Se4]0 clusters (Figure 16D). We have analyzed the spectra of Figures 15 and 16 by using the same procedures as described previously.13 The solid lines drawn through the spectra in these figures are theoretical curves generated from eq 7 with the parameters listed in Table 6. All Mo¨ssbauer spectra of protein-bound Fe3S4 clusters reported thus far have been analyzed with the assumption that the A tensors of the three iron sites are collinear and that their principal axis frames coincide with the frame of the zero-field splitting tensor. Because the latter reflects a global property of the Fe3S4 cluster while the former describes the local sites, it is quite unlikely that this assumption is correct. However, the need to restrict the large number of unknowns (nearly 40 if all tensors of eq 7 have different principal axis frames) makes it impractical to allow the A tensors to occupy different principal axis frames. For applied fields